Actinobacillus pleuropneumoniae LAMP diagnostic kit

A diagnostic kit, pleuropneumonia technology, applied in the field of Porcine Infectious Actinobacillus pleuropneumoniae LAMP diagnostic kit, can solve the problems of dyspnea, slow growth of pigs, high mortality, etc., to achieve convenient use, accurate results, and sensitivity high effect

Inactive Publication Date: 2016-09-21
GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In acute onset, the mortality rate is high, with severe difficulty in breathing, elevated body temperature, loss of appetite, and adhesion between the lung and pleura can be seen in necropsy. In chronic onset, the mortality rate is low, but the pig grows slowly and the feed conversion rate is low

Method used

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  • Actinobacillus pleuropneumoniae LAMP diagnostic kit
  • Actinobacillus pleuropneumoniae LAMP diagnostic kit
  • Actinobacillus pleuropneumoniae LAMP diagnostic kit

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Embodiment 1

[0056] Embodiment 1 of the present invention: Actinobacillus pleuropneumoniae LAMP diagnostic kit for porcine infectious pleuropneumoniae, comprising 250 µL of LAMP diagnostic reaction mixture, Bst DNA polymerase 20µL, 4 primers 40µL each, negative control 20µL, positive control 20µL and ultrapure water 500µL; the sequences of the 4 primers are F3:5 ， -TGGAGCTCCTCGATCTCAAG-3 ， ;B3:5 ， -GCCCCACAATGACGTGTAC-3 ， ;FIP: 5 ， -GCAACGGTCACCAGACTCCCGACAACGGAGTGACCTGTC-3 ， ;BIP: 5 ， -AGAGCAGCACCCTGTAACGTTTACGCTTCTGCATTTTCCCG-3 ， The negative control is ultrapure water, and the positive control is Actinobacillus pleuropneumoniae genomic DNA; the concentration of the primer F3 and the primer B3 is 10 μmol / L; the primer FIP and the primer BIP The concentration is 20µmol / L.

Embodiment 2

[0057] Embodiment 2 of the present invention: Actinobacillus pleuropneumoniae LAMP diagnostic kit for porcine infectious pleuropneumoniae, comprising 250 µL of LAMP diagnostic reaction mixture, Bst DNA polymerase 20µL, 4 primers 40µL each, negative control 20µL, positive control 20µL and ultrapure water 500µL; the sequences of the 4 primers are F3:5 ， -TGGAGCTCCTCGATCTCAAG-3 ， ;B3:5 ， -GCCCCACAATGACGTGTAC-3 ， ;FIP: 5 ， -GCAACGGTCACCAGACTCCCGACAACGGAGTGACCTGTC-3 ， ;BIP: 5 ， -AGAGCAGCACCCTGTAACGTTTACGCTTCTGCATTTTCCCG-3 ， The negative control is ultrapure water, and the positive control is Actinobacillus pleuropneumoniae genomic DNA; the concentration of the primer F3 and the primer B3 is 5 μmol / L; the primer FIP and the primer BIP The concentration is 10µmol / L.

Embodiment 3

[0058] Embodiment 3 of the present invention: Actinobacillus pleuropneumoniae LAMP diagnostic kit for porcine infectious pleuropneumoniae, comprising 250 µL of LAMP diagnostic reaction mixture, Bst DNA polymerase 20µL, 4 primers 40µL each, negative control 20µL, positive control 20µL and ultrapure water 500µL; the sequences of the 4 primers are F3:5 ， -TGGAGCTCCTCGATCTCAAG-3 ， ;B3:5 ， -GCCCCACAATGACGTGTAC-3 ， ;FIP: 5 ， -GCAACGGTCACCAGACTCCCGACAACGGAGTGACCTGTC-3 ， ;BIP: 5 ， -AGAGCAGCACCCTGTAACGTTTACGCTTCTGCATTTTCCCG-3 ， The negative control is ultrapure water, and the positive control is Actinobacillus pleuropneumoniae genomic DNA; the concentration of the primer F3 and the primer B3 is 20 μmol / L; the primer FIP and the primer BIP The concentration is 40µmol / L.

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Abstract

The invention discloses an actinobacillus pleuropneumoniae (APP) LAMP diagnostic kit. According to the invention, a complete LAMP system is constructed; an LAMP amplification method can use a water bath kettle or a heat preservation cup for amplification, the results are visible to naked eyes, the operation is relatively simple, basic on-site quarantine is particularly facilitated, and the sensitivity is high. The kit constructed by a primer designed by the invention can detect APP 1-15 types and has the sensitivity of 0.3 pg/L which is 100 times higher than that of PCR; thus, the established LAMP diagnostic method can perform rapid field detection in pig farms, and is accurate in result and convenient to use.

Description

technical field [0001] The invention relates to the field of livestock breeding and disease prevention and control, in particular to a diagnostic kit for Actinobacillus pleuropneumoniae LAMP. Background technique [0002] Porcine infectious pleuropneumonia is caused by Actinobacillus pleuropneumoniae ( Actinobacillus pleuropneumoniae , APP) infection causes porcine respiratory disease characterized by fibrous pneumonia. In acute onset, the mortality rate is high, with severe dyspnea, elevated body temperature, loss of appetite, and adhesion of the lungs and pleura can be seen at autopsy. In chronic onset, the mortality rate is low, but the pigs grow slowly and the feed conversion rate is low. Since the disease was discovered in my country in 1987, there have been many reports from all over the province, causing serious economic losses to the pig industry. In recent years, APP resistance has become increasingly serious due to the abuse of antibiotics. Contents of the in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/689C12Q1/6844C12Q2531/119C12Q2545/113
Inventor 余波杨莉史开志
Owner GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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